Modification Of Gluten Research Articles

Wheat gluten-based coatings and films: Preparation, properties, and applications.

Effective food packaging that can protect foodstuffs from physical, chemical, and biological damage and maintain freshness and quality is essential to the food industry. Wheat gluten shows promise as food packaging materials due to its edibility, biodegradability, wide availability, low cost, film-forming potential, and high resistance to oxygen. The low mechanical properties and poor water permeability of wheat gluten coatings and films limit their wide applications; however, some inferior properties can be improved through various solutions. This work presents a comprehensive review about wheat gluten-based coatings and films, including their formulation, processing methods, properties, functions, and applications. The mechanical and water resistance properties of coatings and films can be reinforced through wheat gluten modification, combinations of different processing methods, and the incorporation of reinforcing macromolecules, antioxidants, and nanofillers. Antioxidants and antimicrobial agents added to wheat gluten can inhibit microbial growth on foodstuffs, maintain food quality, and extend shelf life. Performances of wheat gluten-based coatings and films can be further improved to expand their applications in food packaging. Current research gaps are identified. Future research is needed to examine the optimal formulation and processing of wheat gluten-based coatings and films and their performance.

A short update on new approaches to celiac disease.

Celiac disease is an autoimmune enteropathy of the small intestine, related to gluten intolerance occurring in genetically predisposed patients. Currently, the only available treatment is a lifelong gluten-free diet. However, the total avoidance of gluten is difficult and poses a challenge to patients, nutritionists and treating physicians. For this reason, scientists have developed in recent years new therapeutic approaches complementary to dietary treatment, such as modification of gluten to make gliadin non-toxic, reduction of the inflammatory response with elafin and Lactococcus Lactis, degradation of gluten by endoproteolytic enzymes, and correction of nutritional deficiencies by adding pseudo-cereals to the diet of celiac patients. This literature review focuses on the different treatment strategies for celiac disease previously studied and summarizes the latest advances in this field.

Characterization of sonicated gluten protein and subsequent rheological properties of model dough and noodles.

The present study aimed to investigate the effects of the thermo-mechanical and rheological properties of a wheat gluten-sonicated model dough and noodles, as well as the effects of ultrasonic frequency (20, 28, 40, 68 and 80 kHz) on the functional properties and structural features of gluten. Water absorption, stability and developmental time, and viscoelastic behavior of gluten-sonicated model dough were all found to be improved. Water absorption, tensile resistance and stretching distance of noodles increased markedly, whereas cooking loss decreased. Ultrasonication at different frequencies also significantly affected gluten structure, including its surface hydrophobicity, micro-network structure, and secondary and tertiary structures. These alterations then caused changes in its functional characteristics. Compared to untreated gluten, sonicated gluten exhibited significantly increased oil and water capacities (8.75-15.26% and 100.65-127.71% higher than the untreated gluten, respectively), foaming and emulsifying properties, and increased solubility (63.46-98.83% higher than control). In addition, these findings indicated that 40 kHz was the likely resonance frequency of the cavitation bubble in the gluten solution. However, sodium dodecyl sulfate-polyacrylamide gel electrophoresis electropherograms revealed that such treatments did not affect the molecular weight of gluten, which was also consistent with its unchanged disulfide bond content. The present study clarified the impact of frequency on the properties of gluten and model dough. The best frequency for modification of gluten was 40 kHz. Collectively, these findings suggest that ultrasonic technology has the potential for use in modifying wheat gluten and commercial noodle processing. © 2022 Society of Chemical Industry.

Effect of chemical structure of selected phenolic acids on the structure of gluten proteins

Effect of overmixing process and structure of selected phenolic acids belonging to hydroxycinnamic and hydroxybenzoic group on the structure of gluten network were analysed with application of FT-Raman Spectroscopy. Modification of gluten by acids resulted in formation of aggregates and unordered structures at the expense of protein stabilizing structures (e.g. β-sheets or β-turns). Supplementation with most of the acids caused reduction in the amount of disulphide bonds in the most stable conformation (g-g-g). Changes in the molecular organization of gluten proteins depended on the chemical structure of particular acids. The presence of bands assigned to aggregates was connected with the number of OH groups present at the aromatic ring of the acids. Acids belonging to hydroxycinnamic group did not incorporate or incorporate only partially into gluten network by formation of covalent or hydrogen bonds. Spectrophotometric analysis showed that hydroxycinnamic acids can interact stronger with gluten proteins compared to hydroxybenzoic acids.

Microbial Transglutaminase Is a Very Frequently Used Food Additive and Is a Potential Inducer of Autoimmune/Neurodegenerative Diseases.

Microbial transglutaminase (mTG) is a heavily used food additive and its industrial transamidated complexes usage is rising rapidly. It was classified as a processing aid and was granted the GRAS (generally recognized as safe) definition, thus escaping full and thorough toxic and safety evaluations. Despite the manufacturers claims, mTG or its cross-linked compounds are immunogenic, pathogenic, proinflammatory, allergenic and toxic, and pose a risk to public health. The enzyme is a member of the transglutaminase family and imitates the posttranslational modification of gluten, by the tissue transglutaminase, which is the autoantigen of celiac disease. The deamidated and transamidated gliadin peptides lose their tolerance and induce the gluten enteropathy. Microbial transglutaminase and its complexes increase intestinal permeability, suppresses enteric protective pathways, enhances microbial growth and gliadin peptide’s epithelial uptake and can transcytose intra-enterocytically to face the sub-epithelial immune cells. The present review updates on the potentially detrimental side effects of mTG, aiming to interest the scientific community, induce food regulatory authorities’ debates on its safety, and protect the public from the mTG unwanted effects.

Viscoelastic properties, antioxidant activities and structure of wheat gluten modified by rice bran

This study presents as a novel rice bran (RB) treatment method for gluten modification. The effects of the method on viscoelastic, and antioxidant properties, as well as the structure of wheat gluten were investigated. RB treatment improved the viscoelastic properties and antioxidant activities (Oxygen radical absorbance capacity and DPPH radical scavenging activity) of modified gluten. Additionally, the modified gluten showed a higher polyphenols content (especially bound polyphenols) and higher percentage of branched chain amino acids compared to control gluten. RB treatment achieved these effect by triggering aggregation of gluten and reaction between amino (-NH2) groups of lysine of gluten proteins and RB polyphenols thereby strengthening its network structure and changing its fluorescence. Moreover, increase in β-turn conformation proved the improvement of gluten aggregation. This study demonstrates that RB treatment modifies the characterization of gluten, and this approach reduces the cost of producing modified gluten and promotes the utilization of RB.

Thermally induced gluten modification observed with rheology and spectroscopies

The protein vital gluten is mainly used for food while interest for non-food applications, like biodegradable materials, increases. In general, the structure and functionality of proteins is highly dependent on thermal treatments during production or modification. This study presents conformational changes and corresponding rheological effects of vital wheat gluten depending on temperature. Dry samples analyzed by X-ray photoelectron spectroscopy (XPS), Fourier-transform infrared spectroscopy (FTIR) and thermalgravimetric analysis coupled with mass spectrometry (TGA-MS) show surface compositions and conformational changes from 25 to 250 °C. Above 170 °C, XPS reveals a decreased N content at the surface while FTIR band characteristics for β-sheets prove structural changes. At 250 °C, protein denaturation accompanied by a significant mass loss due to dehydration and decarbonylation reactions is observed. Oscillatory measurements of optimally hydrated vital gluten describing network properties of the material show two structural changes along a temperature ramp from 25 to 90 °C: at 56–64 °C, the temperature necessary to trigger structural changes increases with the ratio of gliadin to total protein mass, determined by reversed-phase high performance liquid chromatography (RP-HPLC). At a temperature of 79–81 °C, complete protein denaturation occurs. FTIR confirms the denaturation process by showing band shifts with both temperature steps.

Challenges to drug discovery for celiac disease and approaches to overcome them

ABSTRACTIntroduction: The only available effective treatment for celiac disease (CD) is strict and long-term compliance with a gluten-free diet. Dietary gluten restriction must be strict and long term, but is difficult to achieve in many cases and alternative dietary strategies have been investigated in the past few years.Areas covered: This review highlights the progress that has been made in the development of new therapeutics for CD. Detailed information is provided on the targets of drugs for CD as their related mechanisms of action. The therapies are classified in five mechanisms: modification of gluten, intraluminal therapies, immunomodulation, intestinal permeability and modulation of adaptative response. The actual development phase and future approach are also described and discussed.Expert opinion: There are several limitations in each of the treatment targets related either through complications or the lack of complete response to a normal gluten containing diet. It is clear that the most desired therapy for celiac patients would induce gluten tolerance and progress has been made as per the treatments described herein. Therefore, it is shortly expected that curative or complimentary tools to a gluten free diet will be available that will improve the quality of life of CD sufferers.

New coeliac disease treatments and their complications

The only accepted treatment for coeliac disease is strict adherence to a gluten-free diet. This type of diet may give rise to reduced patient quality of life with economic and social repercussions. For this reason, dietary transgressions are common and may elicit intestinal damage. Several treatments aimed at different pathogenic targets of coeliac disease have been developed in recent years: modification of gluten to produce non-immunogenic gluten, endoluminal therapies to degrade gluten in the intestinal lumen, increased gluten tolerance, modulation of intestinal permeability and regulation of the adaptive immune response. This review evaluates these coeliac disease treatment lines that are being researched and the treatments that aim to control disease complications like refractory coeliac disease.

Nuevas terapias en la enfermedad celiaca y sus complicaciones

The only accepted treatment for coeliac disease is strict adherence to a gluten-free diet. This type of diet may give rise to reduced patient quality of life with economic and social repercussions. For this reason, dietary transgressions are common and may elicit intestinal damage. Several treatments aimed at different pathogenic targets of coeliac disease have been developed in recent years: modification of gluten to produce non-immunogenic gluten, endoluminal therapies to degrade gluten in the intestinal lumen, increased gluten tolerance, modulation of intestinal permeability and regulation of the adaptive immune response. This review evaluates these coeliac disease treatment lines that are being researched and the treatments that aim to control disease complications like refractory coeliac disease.

Potential of Lactobacillus plantarum ccm 3627 and Lactobacillus brevis ccm 1815 for fermentation of cereal substrates

Lactobacillus is the most representative strain in a group of lactic acid bacteria, which perform an essential role in the preservation and production of wholesome foods. Lactic acid fermentation is the oldest traditional method for preparation of fermented vegetables, meat products, dairy products and cereal foods. Cereal grains are considered to be one of the most important sources of dietary proteins, carbohydrates, vitamins, minerals and fibre for people. The main exploitation of cereals is to prepare sourdough, which is a mixture of wheat, rye or other cereal flour with water and contains yeasts and lactobacilli. The basic biochemical changes that occur in sourdough bread fermentation are acidification of the dough with organic acids produced by the lactobacilli and leavening with carbon dioxide produced by the yeast and the lactobacilli. Acidification perhaps initiate enzymatic processes of proteins and phytates degradation. Lactobacilli produce various enzymes which make flavour precursors, improve of mineral bioavailability or degrade celiac active peptides, because some species of lactobacilli produce specific peptidases during growth, which are capable to hydrolyze hardly cleavable, celiac-active proline-rich peptides. Microbial fermentation with selected strains of lactobacilli may be new alternative approach for modification of gluten by hydrolysis. In this paper are described growth characteristics and intracellular aminopeptidases activities of Lactobacillus plantarum CCM 3627 and Lactobacillus brevis CCM 1815. Work was focused on characterization of the lactobacilli for potential usage as a starter culture in further fermentation experiments.

Novel approaches for enzymatic gluten degradation to create high-quality gluten-free products

Celiac disease (CD), a chronic enteropathy of the small intestine caused by ingestion of gluten, is one of the most prevalent food hypersensitivities worldwide. The essential treatment is a strict lifelong gluten-free diet based on the avoidance of gluten-containing products from wheat, rye, barley and, in rare cases, oats. Products made from naturally gluten-free raw materials often have inferior nutritional, textural and sensory properties compared to the corresponding gluten-containing products. Therefore, the incorporation of wheat, rye and barley flours after efficient removal of the harmful component gluten into gluten-free products would be beneficial. Gluten modification resulting in decreased CD-immunoreactivity may be achieved via the formation of crosslinks using microbial transglutaminase. To effectively eliminate CD-immunoreactivity, plant, fungal, bacterial, animal or engineered peptidases are capable of degrading gluten proteins and peptides into harmless fragments. The application of peptidases from germinated cereal grains, fungal peptidases and/or lactic acid bacteria during food processing yielded high-quality sourdough wheat breads, pasta, wheat starch and bran, rye products and beer, all with gluten contents below the Codex Alimentarius threshold of 20mg/kg for gluten-free products. As with all gluten-free products, the legislative compliance of such treated materials needs to be monitored closely. Provided that all safety requirements are met, gluten-containing raw materials treated in an adequate way to remove CD-active gluten fragments may be used together with naturally gluten-free ingredients to create an extended choice of high-quality gluten-free products.

Microbial Proteases in Baked Goods: Modification of Gluten and Effects on Immunogenicity and Product Quality.

Gluten-related diseases are a range of inflammatory disorders of the small intestine, characterized by an adverse response to gluten ingestion; therefore, the treatment is a gluten withdrawal. In spite of the increased market of gluten-free products, widely available breads with high acceptability are still missing due to the technological challenge of substituting the special gluten properties. Instead of using alternative ingredients for baking, some attempts have been done to decrease gluten immunogenicity by its enzymatic degradation with microbial proteases. Although the gluten immunogenicity reduction has been reached to an acceptable level, some quality parameters of the products are affected. This review focus on the use of microbial peptidases to prepare less immunogenic baked goods and their effect on product quality.

Gluten degradation in wheat flour with Aspergillus niger prolyl-endopeptidase to prepare a gluten-reduced bread supplemented with an amaranth blend

Celiac disease and other-gluten related disorders are characterized by an inadequate immune reaction against dietary gluten. Some strategies have been developed for gluten modification, to reduce or avoid the immune response. The aim of this study was to modify gluten proteins in wheat flour using the Aspergillus niger prolyl-endopeptidase (AnPEP) to elaborate bread supplemented with amaranth blends, evaluating its immunoreactivity and technological quality. The reaction conditions: enzyme (dilutions 1:50, 1:100 and 1:500), temperature (35 or 40 °C), substrate (WF) (10% or 20%, w/v), time (1–8 h), gluten content by R5-ELISA, and the protein composition by SE-HPLC were analyzed. For bread-making, 60% of modified WF and 40% of amaranth blends (20:80, 30:70, 40:60; raw: popped, respectively), and two fermentation times (52 and 90 min) were tested. Specific volume and gluten content were evaluated. The best reaction conditions were (1:50) AnPEP, 20% WF, and 8 h incubation at 35 °C. The bread supplemented with the 20:80 amaranth blend showed a higher specific volume and after a 52 min fermentation had 99% less immunogenic gluten than the wheat bread. The use of the amaranth blend in combination with WF modified by AnPEP is an effective way to obtain gluten-reduced breads with acceptable quality.

Statement of Retraction

This article refers to:RETRACTED ARTICLE: Prospects of gluten modification to rationalize celiac sprue toxicity – an immunological approach

The present and the future in the diagnosis and management of celiac disease.

Celiac disease is an autoimmune enteropathy caused by gluten in genetically predisposed individuals. In celiac disease, adaptive and innate immune activation results in intestinal damage and a wide range of clinical manifestations. In the past, celiac disease was thought to result in signs and symptoms solely related to the gastrointestinal tract. Now, more than half of the adult population presents with extra-intestinal manifestations that can also be expected to improve on a gluten-free diet. For this reason, it is recommended that physicians have a low threshold of suspicion for celiac disease. Current knowledge of the immune pathogenesis of this autoimmune disease has served as a catalyst for the development of novel diagnostic tools and therapeutics.Over the years, highly sensitive and specific serological assays, in addition to genetic markers, have been found to target specific steps in the cascade pathway of celiac disease. Also the advent of the gluten challenge has enabled experts to design diagnostic algorithms and monitor clinical responses in clinical trials. The gluten challenge has provided substantial benefit in the advance of novel therapeutics as an adjuvant treatment to the gluten free diet. Generally, a strict gluten-free diet is highly burdensome to patients and can be limited in its efficacy. Alternative therapies—including gluten modification, modulation of intestinal permeability and immune response—could be central to the future treatment of celiac disease.

Immunological appraisal of modified gluten to trim down celiac toxicity

The aim of the present research was to modify wheat gluten by binding methionine to gluten proteins to develop bread for celiac disease (CD) patients. The highest protein content, wet gluten content, dry gluten content and sodium dodecyl sulphate-sedimentation value were shown by the wheat variety AARI-11, therefore, it was selected for gluten modification. The bound methionine to gluten proteins was found increasing along the reaction time as the reaction proceeds and at a maximum near to 60 minutes and then it starts decreasing. The lowest immunoreactivity of the modified gluten was obtained near to 60 min of reaction at pH 10. The results for immunoglobulin A (IgA) index showed that the serum of each patient had positive IgA index to gliadins from unmodified gluten, but just sera of two patients had positive IgA index to gliadins from modified gluten and when these proteins were digested, the sera of no patient's serum had positive IgA reactivity. Among physical characteristics of breads 2 hours after baking, the specific volume of the modified gluten containing bread (4.13 ± 0.14 cm3/g) was lower than the control bread (4.59 ± 0.21 cm3/g). However, bread made with modified gluten had higher specific volumes than other gluten-free breads. Texture of the modified gluten was also affected by modification. Finally, the gluten content in the modified gluten bread was 79 ppm which is under the limits set by the Codex Alimentarius for food with reduced gluten content should have from 20 to 100 ppm. The study concludes that the incorporation of steric immensity into gluten proteins in order to shun immune recognition is the most promising approach to acquire wheat-based products that are tolerated by CD patients.

Selective inhibition of the gliadin-specific, cell-mediated immune response by transamidation with microbial transglutaminase

CD is an immune-mediated enteropathy caused by the ingestion of wheat gluten. The modification of gluten by intestinal tTGase plays a crucial role in CD pathogenesis. In this study, we observed that extensive transamidation of wheat flour with K-C2H5 by mTGase yielded spf and K-gliadins fractions. By Western blot, we found that these modifications were associated with strongly reduced immune cross-reactivity. With the use of DQ8 tg mice as a model of gluten sensitivity, we observed a dramatic reduction in IFNγ production in gliadin-specific spleen cells challenged with spf and K-gliadins in vitro (n=12; median values: 813 vs. 29 and 99; control vs. spf and K-gliadins, P=0.012 for spf, and P=0.003 for K-gliadins). For spf, we also observed an increase in the IL-10/IFNγ protein ratio (n=12; median values: 0.3 vs. 4.7; control vs. spf, P=0.005). In intestinal biopsies from CD patients challenged in vitro with gliadins (n=10), we demonstrated further that K-gliadins dramatically reduced the levels of antigen-specific IFNγ mRNA in all specimens responsive to native gliadins (four of 10; P<0.05). As cytotoxic effects have been described for gliadins, we also studied GST and caspase-3 activities using the enterocytic Caco-2 cell line. We found that neither activities were modified by flour transamidation. Our results indicate that K-C2H5 cross-linking via mTGase specifically affects gliadin immunogenicity, reversing the inducible inflammatory response in models of gluten sensitivity without affecting other aspects of the biological activity of gliadins.

Immunological effects of transglutaminase-treated gluten in coeliac disease

Coeliac disease pathogenesis is characterized by an immune response triggered, in genetically predisposed subjects, by ingested gluten and its withdrawal from the diet is the only available therapy. However, enzymatic modification of gluten through the insertion of lysine to avoid antigen presentation could represent a new therapeutical approach for patients. Sixty-six duodenal biopsies from 17 coeliac patients were cultured for 48 h with gluten or enzymatically-modified gluten (treated with human recombinant transglutaminase type 2 or bacterial transglutaminase, with or without lysine). Interferonγ, anti endomisium and anti transglutaminase IgA antibodies, lactate dehydrogenase and transglutaminase activity were measured in the culture medium. Transglutaminase type 2 expression was evaluated on biopsies by immunohistochemistry. Gluten and transglutaminase-treated gluten increased by 13-15 fold interferon γ release, as well as antibodies, transglutaminase activity, and the immunohistochemical expression of transglutaminase type 2. Addition of lysine to the enzymatic modification of gluten normalized interferon γ, antibodies, transglutaminase activity and immunohistochemical expression of transglutaminase type 2. Lactate dehydrogenase did not differ among the studied groups. Enzymatic modification of gluten by transglutaminase plus lysine prevents the immunologic effects on cultured duodenal biopsies from coeliac patients and could be tested as an alternative therapy in coeliac disease.

Celiac Disease: Advances in Treatment via Gluten Modification

Celiac disease (CD) is an autoimmune enteropathy that occurs in genetically susceptible individuals carrying the prerequisite genetic markers HLA DQ2 or DQ8. These genetic markers are present in approximately 30% of the population, and the worldwide prevalence of CD is estimated to be approximately 1%-2%. Currently a gluten-free diet is the only treatment for CD, but novel therapies aimed at gluten modification are underway. This review will discuss gluten-based therapies including wheat alternatives and wheat selection, enzymatic alteration of wheat, oral enzyme supplements, and polymeric binders as exciting new therapies for treatment of CD.